Effects Of Kaempferol, An Active Ingredient Of Buddleja Buddleja, On Hyperosmotic-induced Inflammatory Damage In Human Corneal Epithelial Cells And P38/MAPK Signaling Pathwa

Objective:Aims to explore the effective constituent of Buddleja officinalis,kaempferol(KAE)’s effect on human corneal epithelial cells(HCEC)model induced by the environment of hyperosmolarity and its mechanism.Methods:HCEC was cultured in DMEM/F12 medium,and the morphology and quantity of HCEC were observed under an inverted microscope.CCK8 was used to detect the toxicity and cell viability of KAE,and the best drug concentration was selected.The groups were normal group,hyperosmotic model group,kaempferol group and solvent group.A certain amount of sodium chloride was added into the culture medium to create 450 MOSm hypertonic environment.On this basis,KAE was used to intervene,and the specific mechanism of HCEC after hypertonic treatment and drug action was studied.The proliferation of HCEC was detected by Ki67,the apoptosis of HCEC was detected by TUNEL staining,the protein expression levels of TNF-α,IL-6and P38 were detected by Western Blot,and the m RNA relative expression levels of TNF-α,IL-6 and P38 were detected by fluorescence quantitative PCR.Results:1.Compared with the complete medium,the expressions of TNF-α and IL-6inflammatory factors in HCEC on 450 MOSM medium were significantly increased(P< 0.01),P38 phosphorylation occurred,and p38 MAPK pathway was activated.2.Compared with hyperosmotic group,KAE treatment could significantly reduce the protein expression levels of TNF-α and IL-6(P < 0.01),reduce the relative m RNA expression levels of TNF-α and IL-6(P < 0.01),significantly inhibit cell apoptosis(P< 0.01),and promote cell proliferation(P < 0.01).3.Compared with hypertonic group,KAE treatment could significantly reduce the protein expression level of P38(P < 0.01)and the relative expression level of P38 m RNA(P < 0.01),and P38 signaling pathway was involved in this process.Conclusions:KAE,the effective constituent of Buddleja officinalis,inhibited the production and expression of inflammatory cytokines in the hyperosmotic HCEC model,and the p38 MAPK pathway played an important role.